CN104698183A - Liver cancer tumor marker, antibody and application thereof - Google Patents
Liver cancer tumor marker, antibody and application thereof Download PDFInfo
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- CN104698183A CN104698183A CN201310647697.7A CN201310647697A CN104698183A CN 104698183 A CN104698183 A CN 104698183A CN 201310647697 A CN201310647697 A CN 201310647697A CN 104698183 A CN104698183 A CN 104698183A
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- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
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- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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- G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
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Abstract
The invention relates to a liver cancer tumor marker, an antibody and an application thereof. The liver cancer tumor marker is carbohydrate antigen containing Fuc alpha1,2 structure, the Fuc alpha1,2 structure is alpha1-2 fucoside bond, and is from glycosphingolipid or N-glycosidic bond type glycoprotein. The carbohydrate antigen containing Fuc alpha1,2 structure has special high expression phenomenon in liver cancer tissue, and can be the tumor marker for liver cancer diagnosis. According to the invention, a polyclonal antibody and a monoclonal antibody can be obtained by taking the carbohydrate antigen containing Fuc alpha1,2 structure as immunogen, and the carbohydrate antigen containing Fuc alpha1,2 structure is helpful for liver cancer diagnosis as well as rapidly and accurately finding liver cancer.
Description
Technical field
The present invention relates to tumor markers technical field, particularly relate to a kind of hepatic carcinoma mark, its antibody and application.
Background technology
Liver cancer is one of common malignant tumour, is divided into primary carcinoma of liver and metastatic hepatic carcinoma, and wherein primary carcinoma of liver is the most common and deterioration degree is higher.All about there are 600,000 people's new trouble liver cancer in the world every year, in numerous malignant tumour, ranked fifth position.Wherein, liver cancer accounts for 55% of global onset of liver cancer number at China's patient's number, and this serious threat health and lives safety of our people, its danger should not be underestimated.Therefore, be badly in need of finding a kind of effectively and there is specific tumor markers to help early diagnosis or the anaphase of liver cancer.
Due to the non-sensitive type of chemotherapy of hepatocellular carcinoma radiotherapy, the treatment of liver cancer is made still to lack effective treatment.Therefore, the task of top priority that a kind of therapy target of practical feasibility or new method become Hepatoma therapy is found.Research finds, compound (Glpcokonjugaten) gripped by sugar is the very important biological micromolecule of a class that cell membrane or even albumen exist, and they are for intracellular signaling and maintain normal physiological function and all play a part very crucial.And in tumour, exception often occurs the structure of these molecules, become carbohydrate antigen.These carbohydrate antigen can express the glycoprotein surface at the tumor cell surface of exception or exception, and there is close relationship with the generation of tumour and development.What the carbohydrate antigen of abnormal tumor found at present has T, Tn, β 1, the N-glycan antigen of 6 branches, and as glycosyl sphingolipids such as Lewis a, Lewis b.
This area also needs to find more hepatic carcinoma mark, and prepares corresponding antibody, for the diagnosis fast and accurately of liver cancer.
Summary of the invention
The present inventor finds in liver cancer tissue, have specificity overexpression phenomenon containing the carbohydrate antigen of Fuc α 1,2 structure by research, and does not have this high expressed phenomenon in the normal structure that cancer is other, the present invention is based on this and has found.
The invention provides a kind of hepatic carcinoma mark, its antibody and application, the hepatic carcinoma mark provided clearly can characterize the generation of liver cancer, and its antibody capable provided is enough in the kit preparing diagnosing liver cancer.
The present invention includes following content:
In first aspect, the invention provides a kind of hepatic carcinoma mark, it is carbohydrate antigen, described Fuc α 1,2 structure and α 1 → 2 fucose glycosidic bond containing Fuc α 1,2 structure.
In the present invention, the carbohydrate antigen containing Fuc α 1,2 structure is sometimes referred to as the sugar chain containing Fuc α 1,2 structure hereinafter, and its implication is identical, can replace in the present invention.
As preferably of the present invention, the described carbohydrate antigen containing Fuc α 1,2 structure derives from glycosyl sphingolipid, and namely the described carbohydrate antigen containing Fuc α 1,2 structure is a part for glycosyl sphingolipid.
As preferably of the present invention, the described carbohydrate antigen containing Fuc α 1,2 structure derives from N-glucosides of bonding glycoprotein, and namely the described carbohydrate antigen containing Fuc α 1,2 structure is a part for N-glucosides of bonding glycoprotein.
In second aspect, the invention provides a kind of polyclonal antibody, is produced by the carbohydrate antigen immune animal containing Fuc α 1,2 structure.
As preferably of the present invention, described animal is mouse.
In the third aspect, the invention provides a kind of monoclonal antibody, is that the splenocyte that obtains containing the carbohydrate antigen immune mouse of Fuc α 1,2 structure and mouse lymphoma cell merge the hybridoma obtained and produce.
As preferably of the present invention, described mouse lymphoma cell is P2-X63-Ag8.653.
In fourth aspect, the invention provides a kind of diagnosing cancer of liver kit, comprise the polyclonal antibody described in second aspect or the monoclonal antibody described in the third aspect.
In the 5th, the invention provides the polyclonal antibody described in second aspect or the monoclonal antibody described in the third aspect is preparing the application in diagnosing cancer of liver kit.
In the 6th, the invention provides the polyclonal antibody described in second aspect or the application of the monoclonal antibody described in the third aspect in histogenic immunity dyeing.
Beneficial effect of the present invention is: the present invention finds containing Fuc α 1 by research, the carbohydrate antigen of 2 structures has specificity overexpression phenomenon in liver cancer tissue, and this high expressed phenomenon is not had in the normal structure that cancer is other, judge that the carbohydrate antigen containing Fuc α 1,2 structure can as the tumor marker of diagnosing cancer of liver based on this.The present invention is that immunogene has prepared its polyclonal antibody and monoclonal antibody with the carbohydrate antigen containing Fuc α 1,2 structure, for the diagnosis of liver cancer, contributes to finding liver cancer quickly and accurately.
Accompanying drawing explanation
Fig. 1 is the second order ms figure of the sugar chain containing Fuc α 1,2 structure, and with the sugar chain of Fuc α 1,2 structure of C24 and C16 for example, wherein C24 and C16 refers to the length of the fatty acid chain of ceramide moiety.
Fig. 2 is three grades of mass spectrograms of the sugar chain containing Fuc α 1,2 structure, and with the sugar chain of Fuc α 1,2 structure of C26 and C16 for example, wherein C26 and C16 refers to the length of the fatty acid chain of ceramide moiety.
Fig. 3 is the level Four mass spectrogram of the sugar chain containing Fuc α 1,2 structure, and with the sugar chain of Fuc α 1,2 structure of C26 and C16 for example, wherein C26 and C16 refers to the length of the fatty acid chain of ceramide moiety.
Fig. 4 is the sugar chain content balance result containing Fuc α 1,2 structure in liver cancer tissue and cancer beside organism, and wherein * * represents p<0.001.
Fig. 5 is for containing Fuc α 1, the carbohydrate antigen of 2 structures and the immunostaining results of its monoclonal antibody, wherein arrow instruction positional representation carbohydrate antigen and its monoclonal antibody have combination, there are more carbohydrate antigen and the combination of its monoclonal antibody in result display liver cancer tissue, and in cancer beside organism, have no this combination.
Embodiment
Below in conjunction with embodiment, embodiment of the present invention are described in detail.It will be understood to those of skill in the art that following examples are only the preferred embodiments of the present invention, so that understand the present invention better, thus should not be considered as limiting scope of the present invention.For a person skilled in the art, the present invention can have various modifications and variations, within the spirit and principles in the present invention all, and any amendment done, equivalent replacement or improvement etc., all should be included within protection scope of the present invention.Experimental technique in following embodiment, if no special instructions, is conventional method; Experiment material used, if no special instructions, is and is purchased available from routine biochemistry chemical reagent work.
Embodiment 1
Be separated containing Fuc α 1 from histone or haemocyanin, the sugar chain of 2 structures, concrete steps are as follows: use PNGase F enzyme reagent kit (New England, Britain), the kit instructions provided according to manufacturer, extract albumen N sugar chain, and carry out multi-stage ms analysis to albumen N sugar chain, its process is as follows:
(1) get 500 microgram histones or haemocyanin is placed in centrifuge tube, add 5 microlitre 10 × denaturants, 45 microlitre ultrapure waters, mixing, then boil 10min, then centrifugal.
(2) be cooled to room temperature, add 5 microlitre 10 × G7 damping fluids, 5 microlitre 10%NP-40 damping fluids, add 1 microlitre PNGase F enzyme, spend the night at 37 DEG C.
(3) then dry sample, cross SEP-C18 post, sample, further across after vacuum drying, adds the freshly prepared reducing solution of 300 microlitre under ice bath---NaBH
4solution (10mg/mL in0.01MNaOH), left at room temperature overnight.
(4) under ice bath, add several acetic acid cessation reactions, be then warming up to room temperature, and add 3mL ethanol dry sample.
(5) in order to remove the borate in sample, respectively, in the sample of above-mentioned drying, the following reagent of 3mL is added successively: a, 1% acetic acid: methyl alcohol, b, toluene, c, 1% acetic acid: methyl alcohol, d, toluene, e, 1% acetic acid: methyl alcohol, repeat the step adding reagent-drying, the dry sample finally obtained is the sugar chain cut down from serum antibody.
(6) with the sugar chain that the water-soluble solution of 1mL cuts down.
(7) prepare PGC prepacked column (needing affinity chromatography device), in prepacked column, add following reagent 3mL:a respectively, successively) 1M NaOH; B) HPLC water; C) 80%ACN aqueous solution (including 0.1%TFA); D) 25%ACN aqueous solution (including 0.05%TFA); E) 25%ACN aqueous solution; F) water, carries out washing post; Use 6-8mL water balance pillar again.
(8) to step (7) gained pillar loading step (6) gained sample, 8-10mL water elution pillar is added, to remove impurity and salinity; Then under adding the 25%CAN eluant solution of 3mL, neutral sugar chain (if the sugar of wash-out is acid, needs to add 3m25%ACN aqueous solution (including 0.1%TFA); Vacuum dried sample, adds methyl alcohol sample dissolution, carries out multi-stage ms analysis, and gained mass spectrogram as shown in Figure 1.
(9) by the sugar chain corresponding to peak larger for relative abundance in above-mentioned mass spectrophotometry, multi-stage ms Structural Identification is carried out, to determine the concrete structure of corresponding sugar chain, as shown in Figures 2 and 3.
For containing Fuc α 1, the glycosyl sphingolipid of 2 structures, separation method is as follows: get liver cancer tissue sample or serum sample grinding fragmentation, solubilizer chloroform: methyl alcohol (1:1, v/v) ultrasonic vibration centrifuging and taking supernatant after a hour, quadruplication, add isopropyl alcohol again: hexane: one-level water (55:25:20, v/v/v) ultrasonic vibration centrifuging and taking supernatant after a hour, quadruplication, centrifugal dryer is dry.First by SephadexA25 coagulant liquid dress post, after use chloroform: methyl alcohol: water (30:60:8, v/v/v) solution equilibria, afterwards lipid samples dry in previous step is added chromatographic column, with chloroform: methyl alcohol: water (30:60:8, v/v/v) wash-out obtains neutral glycolipid and collects and obtains neutral glycolipid, and rear centrifugal dryer is dry.First use chloroform: the silica gel column chromatography that No. 60, methyl alcohol (98:2, v/v) solution equilibria, by neutral lipid loading dry in upper step, rear 100mL chloroform: methyl alcohol: water (10:10:2.5, v/v/v) wash-out also collects acquisition neutral glycosphingolipid, nitrogen drying.Methyl alcohol with 0.1mL: neutral glycosphingolipid dry in step in chloroform (1:1, v/v) dissolving, then through mass spectrophotometry.
By mass spectrographic relative quantification, and SPSS software and GraphPad Prism5 software is adopted to carry out Student-Newman-Keuls
a,bcheck analysis proves, containing the sugar chain of Fuc α 1,2 structure high expressed and have statistical significance (Fig. 4) in liver cancer.
Embodiment 2
Specific recognition contains the manufacturing process of the antibody of Fuc α 1,2 structure sugar chain.Buy in known structure containing Fuc α 1, the glycosyl sphingolipid Globo-H(Takara of 2 structures, Japan), get 50 μ g glycosyl sphingolipid Globo-H and combine 0.5mL Freund's complete adjuvant injection mouse, with 50 μ g glycosyl sphingolipid associating 0.5mL incomplete Freund's adjuvants immune mouse again after 2 weeks, after 4 weeks, 400 μ g glycosyl sphingolipids are through peritoneal immunity mouse.Corner of the eyes venous blood sampling after eye socket ,-20 DEG C of frozen serum.Within 4 days, put to death mouse afterwards and get spleen for Fusion of Cells.After three glycosyl sphingolipid immune mouses, conventional ELISA method is adopted to detect at the antibody of the anti-glycosyl sphingolipid of its Serum-induced: to be dissolved in (0.1M NaCO in coating buffer with 15 μ g/mL glycosyl sphingolipids
3, PH9.6,0.1%BSA) 4 DEG C of bags by 96 hole ELISA Plate, close with 5% milk-PBS; Add 1:100 dilute serum successively, the sheep anti-mouse igg of HRP mark, OPD, colour developing stop, and survey OD490nm.The BALB/c mouse of getting non-immune same strain plucks eyeball bloodletting, be placed in EP pipe, its serum does negative control, and de-cervical vertebra puts to death mouse, mouse peritoneal is cut off in superclean bench, draw a little serum-free 1640 nutrient culture media and inject mouse peritoneal, repeatedly blow and beat 4-5 time, sucking-off abdominal cavity inner cell suspension, it is mainly containing macrophage, centrifugal 5 minutes of 1500rpm, supernatant discarded adds HAT nutrient culture media, makes feeder cells suspension and is placed in CO
2cultivate in incubator, stay and do Fusion of Cells use.Mouse P2-X63-Ag8.653(is called for short 653) preparation of lymphoma cell: merge the last fortnight 8-anaguanine (8-AG) and 653 cells are elected cultivation.Merge the same day collect be in exponential phase, 653 cells that motility rate is greater than 95%, with serum free medium RPMI-1640 washed cell once, get 3 × 10
7653 cells for subsequent use.Prepared by mouse boosting cell: select the highest mouse of serum antibody titer, and de-cervical vertebra is put to death, and superclean bench is aseptic gets spleen, grinds and makes single cell suspension, suspends with serum-free 1640 nutrient culture media, collects 2 × 10
8cell is for subsequent use.Fusion of Cells: use conventional method fused cell, 653 cells and mouse boosting cell are merged with the ratio of 1:5 ~ 1:10, serum-free 1640 washs 2-3 time, centrifugal supernatant discarded, gently bottom attack centrifuge tube, that cell precipitation loosens, the 1mL50%PEG of 37 DEG C of preheatings was slowly added dropwise in cell precipitation in 1 minute, limit edged slowly evenly shakes centrifuge tube, drop is dripped along tube wall, leave standstill fusion 90 second, 1mL serum-free 1640 nutrient culture media is added afterwards in 2 minutes, 1mL serum-free 1640 nutrient culture media is added in 1 minute, serum free medium 16401mL is added again in 30 seconds, slowly add serum free medium 164030-40mL again, low-speed centrifugal 6 minutes, abandon supernatant, the calf serum HAT nutrient solution resuspended sedimentation cell gently of 20%, being added dropwise to 96 holes being covered with feeder cells again cultivates in version, every hole 100 μ L.Be put in 37 DEG C, 5%CO
2incubator is cultivated.After to be fused, the cell merged cultivated 14 days in HAT nutrient culture media, uses HAT medium culture after people instead, after waiting two weeks, progressively change the RPMI1640 nutrient culture media of 20% hyclone into.Deng at the bottom of hybridoma confluent cultures version hole 1/4 part time, go culture supernatant, ELISA method detect positive colony.Continuous detecting OD value raises gradually or maintains same level, and its value is greater than negative control 2 times or is judged to be the positive.Retain positive hole, and cloning in time.The previous day of clone prepares 1 × 10 by method noted earlier
5/ ml Turnover of Mouse Peritoneal Macrophages, every hole adds 100 μ L.Select the cell technology in positive hole and clone with limiting dilution assay: getting 500 living cells and be suspended in (50/mL) in 10mL nutrient solution, being inoculated in 96 holes by every hole 100 μ L, being placed on 37 DEG C, 5%CO
2cultivate in incubator, added a nutrient solution in the 5th day, within every three days afterwards, change liquid once.1/3-1/4 antibody test again at the bottom of confluent cultures hole after clone grows up, carries out cloning continuously until the clone's positive rate detected reaches 100%, and the clone strain expansion cultivation selecting positive reaction the strongest is also frozen.Positive monoclonal hybridoma is once proceeded to 24 holes, 6 well culture plates, expand in 1640 complete mediums and cultivate, time bottom cell confluent cultures hole, collect latter 10 minutes 1500rpm centrifugal, discard supernatant, cryopreserving liquid containing the complete culture solution of 10%DMSO dilutes, and is distributed into the cryopreservation tube of 1mL, places for 4 DEG C and places 2 hours in-20 DEG C afterwards for 30 minutes, place 1 hour, be finally transferred in liquid nitrogen frozen for-70 DEG C.The cell clone of a generation, two generations, three generations is frozen in liquid nitrogen all in this way.
Adopt hybridoma cultivation in body in mouse ascites, collect the monoclonal antibody of highly purified anti-Fuc α 1, the 2 structure sugar chain of high-titer.Get the female BAl BIc/c mouse in 6-8 age in week, every mouse peritoneal injects norphytane 0.5mL, one week pneumoretroperitoneum injection hybridoma 1 × 10
7individual/only, 8-10 days afterwards start to collect ascites, ELISA method Positive rate.Centrifuging and taking ascites supernatant also adds isopyknic saturated (NH4)
2sO4 solution mixes and is placed in 40 DEG C of water-bath 4-6 hour, centrifugally abandons supernatant.0.01mol/L PBS (PH=7.4) dissolution precipitation thing, (NH4) is removed in dialysis
2sO4, the Protein G affinity chromatography in accordance with Pharmacia company carries out monoclonal antibody-purified.
The antibody of anti-Fuc α 1,2 structure sugar chain can with prepare diagnosing cancer of liver kit and in dyeing for various histogenic immunity.Example is turned to immune group, get liver cancer patient tumor tissues and make paraffin section, first with 60 DEG C of constant temperature ovens toast 60 minutes, soak 10 minutes in dimethylbenzene again, change a dimethylbenzene and soak 10 minutes again, afterwards respectively successively in absolute ethyl alcohol, 95% ethanol, 80% ethanol, soak 5 minutes in 70% ethanol, PBS carries out antigen retrieval after washing 2-3 time (5 minutes/time), PBS washes 2-3 time (5 minutes/time) again, drip normal serum confining liquid and be placed in room temperature 20 minutes, remove surplus liquid, then add anti-Fuc α 1, the monoclonal antibody 50 μ L of 2 structure sugar chains, leave standstill 37 DEG C 1 hour, PBS washes 5 minutes/time totally 3 times, the sheep anti mouse two dripped again containing the tween-20 of 0.05% resists 50 μ L, place 1 hour in 37 DEG C, PBS washes 5 minutes/time totally 3 times, drip SP to place after 20 minutes in 30 DEG C and wash 5 minutes/time totally 4 times with PBS, 10 minutes are rinsed with PBS after 5-10 minute again with DAB colour developing, haematoxylin redyeing 2 minutes, hydrochloride alcohol breaks up, last tap water dewatered after 15 minutes, transparent, mounting, and microscopy, as shown in Figure 5.Wherein the place of arrow instruction represents that carbohydrate antigen and its monoclonal antibody have combination, more carbohydrate antigen and the combination of its monoclonal antibody is had in result display liver cancer tissue, and in cancer beside organism, have no this combination, prove that the carbohydrate antigen containing Fuc α 1,2 structure is specificity overexpression at liver cancer tissue.
Applicant states, the present invention illustrates detailed features of the present invention and method detailed by above-described embodiment, but the present invention is not limited to above-mentioned detailed features and method detailed, namely do not mean that the present invention must rely on above-mentioned detailed features and method detailed could be implemented.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of auxiliary element, concrete way choice etc. that the present invention selects component, all drops within protection scope of the present invention and open scope.
Claims (10)
1. a hepatic carcinoma mark, it is carbohydrate antigen, described Fuc α 1,2 structure and α 1 → 2 fucose glycosidic bond containing Fuc α 1,2 structure.
2. hepatic carcinoma mark according to claim 1, is characterized in that, the described carbohydrate antigen containing Fuc α 1,2 structure derives from glycosyl sphingolipid.
3. hepatic carcinoma mark according to claim 1, is characterized in that, the described carbohydrate antigen containing Fuc α 1,2 structure derives from N-glucosides of bonding glycoprotein.
4. a polyclonal antibody is produced by the carbohydrate antigen immune animal containing Fuc α 1,2 structure.
5. polyclonal antibody according to claim 4, is characterized in that, described animal is mouse.
6. a monoclonal antibody is that the splenocyte that obtains containing the carbohydrate antigen immune mouse of Fuc α 1,2 structure and mouse lymphoma cell merge the hybridoma obtained and produce.
7. monoclonal antibody according to claim 6, is characterized in that, described mouse lymphoma cell is P2-X63-Ag8.653.
8. a diagnosing cancer of liver kit, comprises the polyclonal antibody as described in claim 4 or 5 or monoclonal antibody as claimed in claims 6 or 7.
9. the polyclonal antibody as described in claim 4 or 5 or monoclonal antibody are as claimed in claims 6 or 7 preparing the application in diagnosing cancer of liver kit.
10. the polyclonal antibody as described in claim 4 or 5 or the application of monoclonal antibody as claimed in claims 6 or 7 in histogenic immunity dyeing.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101779128A (en) * | 2007-06-14 | 2010-07-14 | 弗拉芒区生物技术研究所 | Diagnostic test for the detection of early stage liver cancer |
| CN102066940A (en) * | 2008-06-16 | 2011-05-18 | 中央研究院 | Cancer diagnosis based on levels of antibodies against Globo H and its fragments |
-
2013
- 2013-12-04 CN CN201310647697.7A patent/CN104698183A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101779128A (en) * | 2007-06-14 | 2010-07-14 | 弗拉芒区生物技术研究所 | Diagnostic test for the detection of early stage liver cancer |
| CN102066940A (en) * | 2008-06-16 | 2011-05-18 | 中央研究院 | Cancer diagnosis based on levels of antibodies against Globo H and its fragments |
Non-Patent Citations (4)
| Title |
|---|
| P.F. SPITALNIK ET AL: "THE GLYCOSPHINGOLIPID COMPOSITION OF THE HUMAN HEPATOMA CELL LINE, HEP-G2", 《ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS》 * |
| WU LIHUI ET AL: "Fucosylated Glycans Assosicated with Development and Metastasis of Hepatocellular Carcinoma Cells", 《PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS》 * |
| 朱健: "鞘糖脂组学方法研究肝癌特征性鞘糖脂", 《中国优秀硕士学位论文全文数据库（电子期刊）医药卫生科技辑》 * |
| 王镜岩等: "《生物化学（第三版）》", 30 September 2002 * |
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